Apparatus for generating gas



April 19, 1966 J. H. BREWER APPARATUS FOR GENERATING GAS 2 Sheets-Sheet1 Filed March 21, 1965 INVENTOR. JO/l/V H. EAEWE/P ATTO/FNE'KS April 19,1966 BREWER 3,246,959

APPARATUS FOR GENERATING GAS Filed March 21, 1963 2 Sheets-Sheet 2 JO/M/H. BREWER United States Patent 3,246,959 APPARATUS FOR GENERATING GASJohn H. Brewer, 425 Oak Lane, Towson, Md. Filed Mar. 21, 1963, Ser. No.267,040 3 Claims. c1. 23-282) This invention relates to a gas-producingdevice and method which provides a non-toxic atmosphere for culturingmicroorganisms and more particularly to a device and method which evolvegas for aerobiosis and anaerobiosis under controlled conditions in whicha predetermined volume of gas is evolved.

The growth of many microorganisms is hindered when exposed toatmospheric conditions. Thus, a special apparatus must be supplied fortheir growth and provision of a non-toxic atmosphere must be introducedin the apparatus to sustain the growth of the microorganisms.Microorganisms such as gonococcus, meningococcus and brucella requiresubstantially higher concentration of carbon dioxide over that ofordinary atmosphere for proper growth. Microorganisms that are obligateanaerobes such as the bacilli of tetanus, gas gangrene, botulinus andbacteroides require the absence of oxygen for proper growth. Therefore,non-toxic atmospheres must be provided in the culturing apparatus tomaintain proper growth of the desired microorganisms. The devices andmethods heretofore used are generally inconvenient, expensive andrequire skilled technicians to perform them.

There are many difi'iculties and disadvantges presented when usingaerobic and anaerobic culturing apparatus for use in culturingmicroorganisms. Methods employed for supplying a non-toxic atmosphere inan apparatus for culturing microorganisms are the use of a storedcylinder gas, such as hydrogen, illuminating gas, carbon dioxide or thelike, and chemical adsorbers of oxygen. The cylinder gas is generallystored under pressure in which monometers, gauges, reducing valves areneeded to introduce a measured amount of gas into the apparatus.However, this method has proved inconvenient and expensive-and at timeshighly dangerous because an excess of gas placed into the apparatuscould cause an explosion due to excessive pressure. The necessity ofbulky and expensive component parts of the apparatus does not lenditself to mobility for use outside the laboratory. Other methods alsoemployed, such as chemicals or catalysts which burn out trace amounts ofoxygen remaining in the apparatus are inconvenient, in that skilledtechnicians are needed to handle the chemicals such as red phosphoruswhich is highly comustible in the presence of oxygen.

It is therefore an object of my invention to provide a device in which apredetermined amount of non-toxic gas may be supplied to microaerophilicand anaerobic culturing apparatus, in which the device is disposable,which eliminates the need for gauges, reducing valves, monometers,vacuum pumps, cylinders of gas and'other sources of combustible gas.Also, the danger of explosion from gas under high pressure issubstantially eliminated.

Another object of my invention is to provide a mobile microaerophilicand anaerobic culturing apparatus for use outside the laboratory inwhich the use of highly skilled technicians is not needed; also, toprovide a device as a source of non-toxic gas for aerobiosis andanaerobiosis in which the device is disposable and inexpensive tomanufacture.

My invention generally contemplates a device which is disposable andwhich is capable of evolving a nontoxic atmosphere for culturing aerobicand anaerobic microorganisms in culturing apparatus of the type ingeneral use.

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More specifically, my invention contemplates the provision of a sealedenvelope having a plurality of chambers or compartments. In one of thechambers I provide material which when mixed with a liquid evolves orgenerates a non-toxic gas or atmosphere suitable for aerobic andanaerobic microorganisms. Another compartment receives a liquid andtransfers it at a controlled, relatively slow rate to the firstcompartment whereby the gas or atmosphere is evolved at a similarcontrolled slow rate.

Other objects of my invention will become more apparent from theaccompanying drawings by way of illustration thereof, in which:

FIG. 1 is an elevational view of a gas producing device embodying myinvention;

FIG. 2 is an elevational view in cross-section of an anaerobe jar havingmy improved gas producing device therein;

FIG. 3 is a horizontal sectional view taken along line 33 of FIG. 2;

FIG. 4 is a horizontal sectional view taken along line 4-4 of FIG. 1;

FIG. 5 is an enlarged vertical sectional view taken along line 5--5 ofFIG. 1;

FIG. 6 is an enlarged vertical sectional view taken on line 66 of FIG.1;

FIG. 7 is an elevational view of another embodiment of my invention; and

FIG. 8 is a vertical sectional view along line 8-8 of FIG. 7. 7

My improved apparatus comprises an envelope 10 made of a suitablematerial which is impervious to the atmosphere and to moisture and whichis inert thereto and is also inert to the materials 12 contained in theenvelope and the gas generated thereby. For this purpose I may employ ametallic foil, such as aluminum foil coated on its inner surface with athermoplastic material, such as polyethylene or a polymer or copolymerof vinyl chloride. The envelope 10 may be formed from two panels 14 and15 suitably secured together around their edges 16, 17, 18 and 19 as byheat sealing. The envelope 10 is divided internally into a plurality ofcompartments including a first liquid receiving compartment 20 and asecond gas generating compartment 21. The compartments 20 and 21 areseparated by a suitable partition 22 which is formed by means such asheat sealing and extends diagonally across the envelope. The partition22 includes a liquid transfer portion 23 for transferring liquid at acontrolled slow rate of speed from the first compartment 20 to thesecond compartment 21. For this purpose I prefer to provide porouswick-like means having a porosity which prevents the free flow of liquidtherethrough but through which the liquid may diffuse from the firstcompartment 20 to the second compartment 21. Thus, the partition 22 maybe formed by a heat seal line extending diagonally across envelope 10.Included between the two panels 14 and 15 along the heat seal line 22are strips of a porous material 23 and 24, such as filter paper,blotting paper, cotton twill, sintered glass or the like, which serve asa means to transfer liquid from the first compartment 20 to the secondcompartment 21 and serves to transfer the gas evolved in the secondcompartment '21 in an outward direction.

In the second chamber I provide a suitable gas generating material 12which Will generate or evolve gas when mixed with the liquid from thefirst compartment 20. It is obvious that materials and the proportionsused will vary depending upon the type of gas to be generated and thevolume that is necessary to produce aerobiosis or anaerobiosis. Sincethe apparatus generally used has a volume of approximately 2 liters thegas evolving materials 12 employed by me are calculated to yield avolume of gas of approximately 2 liters at standard temperature andpressure.

For the production of hydrogen the following may be used:

Example 1 Gms. Magnesium turnings 3.5 Sodium chloride 2.5 Zinc chloride1.0

Approximately 10.0 cc. of water previously supplied to the liquidreceiving chamber is transferred at a slow rate to the gas generatingchamber to thereby slowly generate the hydrogen. 1 have found that theuse of sodium chloride accelerates the reaction at a controlled rate andgoes to completion within approximately 30 minutes, but when sodiumchloride is not used thetreaction proceeds at a much slower rate.

Example 2 Sodium borohydride 0.9 gms.

The same procedure is followed as described in Example 1 when addingapproximately 10.0 cc. of water. To accelerate the reaction at acontrolled rate I may use .2 gms. of nickel or cobalt as a catalyst.Also, to prevent the decomposition of the sodium borohydride a watersoluble coating, such as gelatin is provided- Example 3 For theproduction of carbon dioxide:

Gms.

Citric acid 0.7 Sodium carbonate 0.867

Example 4 For the production of acetylene gas:

Calcium carbide 2.95 gms.

The same procedure is followed as described in Example 1 when addingapproximately 10.0 cc. of water. The calcium carbide is in finelydivided form and is treated with an oil to prevent slaking of thecalcium carbide when in contact with air. Acetylene will be generatedupon contacting the calcium carbide with water, however, in order tocontrol the rate of reaction, a watersoluble coating such as gelatin ormethylcellulose, is provided to encase the carbide. Also, the controlledrate of transfer of water from the first compartment 20 to the secondcompartment 21 Controls the evolution of acetylene.

The above examples are illustrative of various types of gas that may beevolved from my device by reacting a liquid with a predetermined amountof gas evolving materials to obtain a predetermined volume of gas.

When using my envelope 10, the liquid receiving compartment 20 isarranged so that it may be readily opened for the introduction of liquidtherein. As shown in FIGS. 1 and 2, a corner 26 is torn or cut away sothat 10.0 cc. of water may be introduced into compartment 20. The waterflows through the liquid transfer portion 23 at a controlled relativelyslow rate into compartment 21 and mixes with materials 12 as previouslydescribed by way of Examples 1 to 4. The gas evolved passes fromcompartment 21 in an outward direction through transfer portion 24.

When using my envelope 10 in accordance with FIGS. 7 and 8 a measuredquantity of liquid is provided in chamber 27. A frangible partition 28is formed by means such as heat sealing panels 14 and 15 of envelope 10as shown in FIG. 7. Also, the measured quantity of liquid may be placedin a frangible envelope and placed in compartment 20. It should be notedthat partition 28 or said frangible envelope is such that it willrupture under slight pressure, yet the pressure will not rupturepartition 22 or the sealed edges 16, 17, 18 and 19 of envelope 10. Afterthe liquid has been transferred into compartment 20 edge 26 is cut ortorn away as shown in FIG. 2. The same procedure is followed aspreviously described from the point in which the liquid is introducedinto compartment 20.

In carrying out an embodiment of my invention reference is had to FIG. 2in which an anaerobe jar is illustrated embodying my invention inoperative form. The jar comprises a container portion 30 which has aflanged rim 31 so that the top 32 may be sealed thereon. Petri dishes 35or culture tubes containing the anaerobic cultures are placed incontainer 30, as shown in FIG. 2. Corner 26 of envelope 10 is cut orsevered therefrom and panels 14 and 15 are spread open so thatcompartment 20 is exposed and a measured amount of liquid, such aswater, is placed therein. The water when placed in compartment 20remains therein for a short period of time enabling the jar to be sealedso that the gas evolved will not escape to the atmosphere. The wickmeans 23 provides suflicient delay time in which the transfer of waterfrom the compartment 20 to compartment 21 permits the jar to be sealedso that the gas evolved is completely contained in the anaerobe jar. Thegas evolved passes from compartment 21 through wick means 24, throughcompartment 20 and finally fills the anaerobe jar. The lid 32 comprisesa clamp 36 which is detachably connected to rim 31 of container 30 andis fastened in sealing engagement with the surface of rim 31 to preventpassage of gas out of the jar. Lid 32 is fitted with a heating element37 sealed in a solid brass tube 38 and has a platinized wire gauze 39secured at opposite ends of lid 32. The oxygen and other gases combinein space 40 when an electrical current is connected to the brass tube 37for a period of 30 minutes. Thus, it will be seen after the oxygen hasbeen completely reacted the atmosphere remaining in the jar is anon-toxic atmosphere which will permit the culturing of anaerobicbacteria. The gas evolved is chemically generated by materials 12 whenreacted with water to give a predetermined volume of gas. My invention,therefore, provides a source of gas Without the use of monometers,gauges, vacuum pumps or cylinders of gas. As is obvious, the procedurefor providing a non-toxic atmosphere for aerobic organisms will vary sothat the oxygen is not chemically reacted but remains as part of thenon-toxic atmosphere.

While I have described the embodiments of my invention in thespecification and shown my device by the illustrated drawings, changesmay be made thereto without departing from the scope of the appendedclaims.

I claim:

1. A device providing a non-toxic atmosphere for use in culturinganaerobic microorganisms comprising: an envelope made of a materialinert to the contents thereof and to the gas generated therein andunaffected by the heat of reaction of the hereinafter mentionedgas-generating material and liquid, said envelope having a firstliquid-receiving chamber and a second gas-generating chamber andmaterial disposed in said second chamber for generating a gaseousatmosphere suitable for culturing anerobic microorganisms when mixedwith a liquid, said envelope being provided'with a partition separatingsaid chambers from each other and having porous fluid transfer meansincluding a water inlet portion and a gas egress portion through whichthe liquid slowly diffuses from the first chamber to the second chamberat a controlled, relatively slow rate to react with the material togenerate gas at a controlled, relatively slow rate and through which thegas which is evolved in the second chamber may dilfuse in an outwarddirection, said first chamber being initially free from liquid adjacentthe porous transfer means and having a frangible portion which may beruptured to afford access for the liquid to the chamber.

2. A device providing a non-toxic atmosphere for 10 use in culturinganerobic microorganisms as set forth in claim 1 in which the partitionbetween the chambers is diagonally disposed and the porous transfermeans comprises a wick extending through the lower portion of thepartition for transferring liquid from the first to the second chamberand a wick near the upper portion of the partition through which the gasmay evolve.

3. A device providing a non-toxic atmosphere for use in culturinganaerobic microorganisms as set forth in claim 1 in which the envelopeis made of an outer layer of metallic foil and an inner layer ofthermoplastic material heat sealed around the edges and across thepartition.

References Cited by the Examiner UNITED STATES PATENTS 2,828,245 3/1958Freaney 195-109 2,981,660 4/1961 Achorn et a1. 195-142 3,013,950 12/1961Gavin 195-142 3,041,250 6/1962 Wolnak et al. 195109 3,102,082 8/1963Brewer 195-139 MORRIS O. WOLK, Primary Examiner.

15 A. LOUIS MONACELL, Examiner.

A. E. TANENHOTTZ, I. H. TA-Y MAN, 111.,

Assistant Examiners.

1. A DEVICE PROVIDING A NON-TOXIC ATMOSPHERIC FOR USE IN CULTURINGANAEROBIC MICROORGANISMS COMPRISING: AN ENVELOPE MADE OF A MATERIALINERT TO THE CONTENTS THEREOF AND TO THE GAS GENERATED THEREIN ANDUNAFFECTED BY THE HEAT OF REACTION OF THE HEREINAFTER MENTIONEDGAS-GENERATING MATERIAL AND LIQUID, SAID ENVELOPE HAVING A FIRSTLIQUID-RECEIVING CHAMBER AND A SECOND GAS-GENERATING CHAMBER ANDMATERIAL DISPOSED IN SAID SECOND CHAMBER FOR GENERATING A GASEOUSATMOSPHERE SUITABLE FOR CULTURING ANEROBIC MICROORGANISMS WHEN MIXEDWITH A LIQUID, SAID ENVELOPE BEING PROVIDED WITH A PARTITION SEPARATINGSAID CHAMBERS FROM EACH OTHER AND HAVING POROUS FLUID TRANSFER MEANSINCLUDING A WATER INLET PORTION AND A GAS EGRESS PORTION THROUGH WHICHTHE LIQUID SLOWLY DIFFUSES FROM THE FIRST CHAMBER TO THE SECOND CHAMBERAT A CONTROLLED, RELATIVELY SLOW RATE TO REACT WITH THE MATERIAL TOGENERATE GAS AT A CONTROLED, RELATIVELY SLOW RATE AND THROUGH WHICH THEGAS WHICH IS EVOLVED IN THE SECOND CHAMBER MAY DIFFUSE IN AN OUTWARDDIRECTION, SAID FIRST CHAMBER BEING INITIALLY FREE FROM LIQUID ADJACENTTHE POROUS TRANSFER MEANS AND HAVING A FRANGIBLE PORTION WHICH MAY BERUPTURED TO AFFORD ACCROSS FOR THE LIQUID TO THE CHAMBER.